JP7706475B2 - Combinations Containing Tricyclic Compounds and Their Use in the Manufacture of Medicaments for the Treatment of HBV - Google Patents

Description

This application claims priority to Chinese patent application CN202010412760.9, filed on May 15, 2020, and Chinese patent application CN202011633373.4, filed on December 31, 2020. This application cites the above Chinese patent applications in their entirety.

[Technical field]
The present invention relates to a pharmaceutical combination, specifically a combination comprising a compound having a tricyclic structure or a pharma- ceutically acceptable salt thereof, and optionally one or two of a hepatitis B surface antigen inhibitor and/or a reverse transcriptase inhibitor, and the use of said combination in the manufacture of a medicament for the treatment of hepatitis B. Specifically disclosed is a combination comprising a compound of formula (I) or a pharma- ceutically acceptable salt thereof, and optionally one or two of a hepatitis B surface antigen inhibitor and/or a reverse transcriptase inhibitor, and the use of said combination in the manufacture of a medicament for the treatment of hepatitis B.

Hepatitis B is an inflammatory response caused by the invasion of the hepatitis B virus, which can lead to a series of problems such as liver pain, liver and spleen enlargement, liver fibrosis, and in severe cases, cirrhosis or even liver cancer. According to statistics, there are approximately 350 to 400 million hepatitis B virus carriers in the world, one-third of which are in China, where 500,000 people die from hepatitis B every year.

At present, there is no specific drug for hepatitis B in the world, and the first-choice drugs for hepatitis B treatment in China are mainly nucleoside drugs, interferon, and traditional Chinese medicines, but they have problems such as high cost and inability to cure, so the development of new anti-hepatitis B drugs is essential. Existing drug monotherapy has problems of drug resistance and not ideal therapeutic effect, so the combination of hepatitis B treatment drugs, especially the combination of hepatitis B drugs with different mechanisms, tends to become a research hotspot and the direction of clinical use.

Patent WO2018153285A1 discloses a compound of formula (I) which is a hepatitis B core protein inhibitor and its use; WO2018214875A1 and WO2018161960A1 disclose a compound of formula (II) and a compound of formula (III) which are hepatitis B surface antigen inhibitors and their use, respectively. Nucleoside or nucleotide reverse transcriptase inhibitors currently clinically recommended include entecavir, tenofovir disoproxil fumarate, etc.

International Publication No. 2018153285 International Publication No. 2018214875 International Publication No. 2018161960

The present invention aims to achieve the goal of treating HBV synergistically through the combination and use of drugs with different mechanisms.

ここで、式（Ｉ）において、
Ｌ_１は単結合又はＣ_１－６アルキルであり、
Ｒ_１はＨ、Ｃｌ、Ｆ、Ｂｒ、Ｉ、或いは任意選択で１、２又は３個のＲ_ａで置換されたＣ_１－３アルキルであり、
環Ａは、４～８員ヘテロシクロアルキル又はＣ_３－８シクロアルキルであり、
Ｒ_２はＨ及びＣ_１－３アルキルであり、
Ｒ_３はＨ及びＣ_１－３アルキルであり、
Ｒ_ａはそれぞれ独立してＨ、Ｆ、Ｃｌ、Ｂｒ、Ｉ、ＮＨ_２、ＯＨ、ＣＮ、Ｃ_１－６アルキル又はＣ_１－６ヘテロアルキル、或いは任意選択で１、２又は３個のＲ’で置換されたＣ_１－６アルキル及びＣ_１－６ヘテロアルキルであり、
Ｒ’はそれぞれ独立してＣｌ、Ｆ、Ｂｒ、Ｉ、ＮＨ_２、ＣＨ_３、ＣＮ及び－Ｎ（ＣＨ_３）_２から選択され、
前記４～８員ヘテロシクロアルキル及びＣ_１－６ヘテロアルキルはそれぞれ、１、２、３又は４個の独立して－Ｏ－、－ＮＨ－、－Ｓ－及びＮから選択されるヘテロ原子又はヘテロ原子団を含む。 The present invention discloses a pharmaceutical combination, said combination being a compound of formula (I) or a pharma- ceutically acceptable salt thereof and any of the following groups a to c:
a. Hepatitis B surface antigen inhibitors;
b. reverse transcriptase inhibitors;
c, Hepatitis B surface antigen inhibitors and reverse transcriptase inhibitors.

Here, in formula (I),
_L1 is a single bond or _C1-6 alkyl;
R ₁ is H, Cl, F, Br, I, or C _1-3 alkyl optionally substituted with 1, 2 or 3 R _a ;
Ring A is a 4-8 membered heterocycloalkyl or C _3-8 cycloalkyl;
_R2 is H and _C1-3 alkyl;
_R3 is H and _C1-3 alkyl;
each R _a is independently H, F, Cl, Br, I, NH ₂ , OH, CN, C _1-6 alkyl or C _1-6 heteroalkyl, or C _1-6 alkyl and C _1-6 heteroalkyl optionally substituted with 1, 2 or 3 R';
R' is independently selected from Cl, F, Br, I, _NH2 , _CH3 , CN, and -N( _CH3 ) ₂ ;
The 4 to 8 membered heterocycloalkyl and C _1-6 heteroalkyl each contain 1, 2, 3 or 4 heteroatoms or heteroatomic groups independently selected from -O-, -NH-, -S- and N.

Preferably,
Here, each R _a is independently H, F, Cl, Br, I, NH ₂ , OH, or CN.

Here, ring A is a 5- to 6-membered heterocycloalkyl.

Wherein ring A is tetrahydrofuranyl, tetrahydropyranyl or dioxanyl.

である。 Here, ring A is

It is.

wherein R ₁ is H, Cl, F, Br, I, or CH ₃ optionally substituted with 1, 2 or 3 R _a . More preferably, wherein R ₁ is H, Cl or CH ₃ .

Here, _R2 is H or _CH3 .

_R3 is H or _CH3 .

Here, L ₁ is —CH ₂ — or —CH ₂ CH ₂ —.

ここで、Ｒ_１、Ｒ_２、Ｒ_３及びＬ_１の定義は、上記と同じである。 In one embodiment, the compound of formula (I) has the structure of formula (I-1):

Here, the definitions of R ₁ , R ₂ , R ₃ and L ₁ are the same as above.

Particularly preferred compounds of formula (I) of the present invention are selected from the specific compounds of the following formulae:

ここで、
Ｒ_１はＨ、ＯＨ、ＣＮ、ＮＨ_２から選択され、或いは任意選択で１、２又は３個のＲで置換されたＣ_１－６アルキル、Ｃ_１－６ヘテロアルキル、Ｃ_２－５アルケニル、Ｃ_２－５ヘテロアルケニル、Ｃ_３－６シクロアルキル又は３～６員ヘテロシクロアルキルから選択される。
Ｒ_２はＨ、ＯＨ、ＣＮ、ＮＨ_２、ハロゲンから選択され、或いは任意選択で１、２又は３個のＲで置換されたＣ_１－３アルキル、Ｃ_１－３ヘテロアルキル、Ｃ_３－６シクロアルキル又は３～６員ヘテロシクロアルキルから選択され、
Ｒ_３は任意選択で１、２又は３個のＲで置換されたＣ_１－６アルキル、Ｃ_３－６シクロアルキルから選択され、
ｍは、０、１、２、３、４又は５から選択され、
ｍが０の場合、Ｒ_１はＯＨ、ＣＮ、ＮＨ_２から選択されなく、
ＲはＨ、ハロゲン、ＯＨ、ＣＮ、ＮＨ_２から選択され、或いは任意選択で１、２又は３個のＲ’で置換されたＣ_１－３アルキル、Ｃ_１－３ヘテロアルキルから選択され、
Ｒ’はＦ、Ｃｌ、Ｂｒ、Ｉ、ＯＨ、ＣＮ、ＮＨ_２、ＣＨ_３、ＣＨ_３ＣＨ_２、ＣＨ_３Ｏ、ＣＦ_３、ＣＨＦ_２、ＣＨ_２Ｆから選択され、
「ヘテロ」は、ヘテロ原子又はヘテロ原子団を表し、前記Ｃ_１－６ヘテロアルキル、Ｃ_２－５ヘテロアルケニル、３～６員ヘテロシクロアルキル、Ｃ_１－３ヘテロアルキルの「ヘテロ」は、それぞれ独立して－Ｃ（＝Ｏ）Ｎ（Ｒ）－、－Ｎ（Ｒ）－、－Ｃ（＝ＮＲ）－、－（Ｒ）Ｃ＝Ｎ－、－Ｓ（＝Ｏ）_２Ｎ（Ｒ）－、－Ｓ（＝Ｏ）Ｎ（Ｒ）－、Ｎ、－Ｏ－、－Ｓ－、＝Ｏ、＝Ｓ、－Ｃ（＝Ｏ）Ｏ－、－Ｃ（＝Ｏ）－、－Ｃ（＝Ｓ）－、－Ｓ（＝Ｏ）－、－Ｓ（＝Ｏ）_２－、－Ｎ（Ｒ）Ｃ（＝Ｏ）Ｎ（Ｒ）－から選択され、
上記の場合のいずれにおいても、ヘテロ原子又はヘテロ原子団の数は、それぞれ独立して１、２又は３から選択される。 In some aspects of the invention, in the above combinations, the Hepatitis B surface antigen inhibitor is selected from one of the compounds of formula (II) or a pharma- ceutically acceptable salt thereof.

Where:
R ₁ is selected from H, OH, CN, NH ₂ , or selected from C _1-6 alkyl, C _1-6 heteroalkyl, C _2-5 alkenyl, C _2-5 heteroalkenyl, C _3-6 cycloalkyl or 3- to 6-membered heterocycloalkyl optionally substituted with 1, 2 or 3 R.
R ₂ is selected from H, OH, CN, NH ₂ , halogen, or is selected from C _1-3 alkyl, C _1-3 heteroalkyl, C _3-6 cycloalkyl or 3- to 6-membered heterocycloalkyl optionally substituted with 1, 2 or 3 R;
R ₃ is selected from C _1-6 alkyl, C _3-6 cycloalkyl optionally substituted with 1, 2 or 3 R;
m is selected from 0, 1, 2, 3, 4 or 5;
When m is 0, _R1 is not selected from OH, CN, _NH2 ;
R is selected from H, halogen, OH, CN, _NH2 , or _C1-3 alkyl, _C1-3 heteroalkyl optionally substituted with 1, 2 or 3 R';
R' is selected from F, Cl, Br, I, OH, CN, _NH2 , _CH3 , _CH3CH2 , _CH3O , _CF3 , _CHF2 , _{CH2F ;}
"Hetero" represents a heteroatom or a heteroatom group, and the "hetero" in the C _1-6 heteroalkyl, C _2-5 heteroalkenyl, 3- to 6-membered heterocycloalkyl, and C _1-3 heteroalkyl is each independently selected from -C(=O)N(R)-, -N(R)-, -C(=NR)-, -(R)C=N-, -S(=O) ₂ N(R)-, -S(=O)N(R)-, N, -O-, -S-, =O, =S, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) ₂ -, and -N(R)C(=O)N(R)-;
In any of the above cases, the number of heteroatoms or heteroatom groups is each independently selected from 1, 2 or 3.

In some embodiments of the invention, in the above combination, the compound of formula (II) is selected from one of the compounds of the following formulae:

In some embodiments of the invention, in the above combination, the compound of formula (II) is selected from one of the compounds of the following formulae:

好ましくは、前記Ｂ型肝炎表面抗原阻害剤は、下記の式（ＩＩｂ）で表される構造を有する。

Preferably, the Hepatitis B surface antigen inhibitor has a structure represented by formula (IIa):

Preferably, the Hepatitis B surface antigen inhibitor has a structure represented by formula (IIb):

ここで、
Ｒ_１はＨ、ＯＨ、ＣＮ、ＮＨ_２から選択され、或いは任意選択で１、２又は３個のＲで置換されたＣ_１－５アルキル、Ｃ_１－５ヘテロアルキル、Ｃ_２－５アルキニル、Ｃ_３－６シクロアルキル及び３～６員ヘテロシクロアルキルから選択され、
Ｒ_２はＨ、ハロゲンから選択され、或いは任意選択で１、２又は３個のＲで置換されたＣ_１－３アルキル及びＣ_１－３ヘテロアルキルから選択され、
ｍは、０、１、２、３、４及び５から選択され、
Ａは任意選択で１、２又は３個のＲで置換されたフェニル又は５～６員ヘテロアリールから選択され、
ＲはＨ、ハロゲン、ＯＨ、ＣＮ、ＮＨ_２、＝Ｏ、ＣＨ_３、ＣＨ_３ＣＨ_２、ＣＨ_３Ｏ、ＣＦ_３、ＣＨＦ_２、ＣＨ_２Ｆから選択され、
前記Ｃ_１－５ヘテロアルキル、３～６員ヘテロシクロアルキル、Ｃ_１－３ヘテロアルキル、５～６員ヘテロアリールの「ヘテロ」は、それぞれ独立してＮ、－Ｏ－、＝Ｏ、－Ｓ－、－ＮＨ－、－（Ｃ＝Ｏ）－、－（Ｓ＝Ｏ）－、－（Ｓ＝Ｏ）_２－から選択され、
上記の場合のいずれにおいても、ヘテロ原子又はヘテロ原子団の数は、それぞれ独立して１、２又は３から選択される。 In another embodiment, the Hepatitis B surface antigen inhibitor is selected from one of the compounds of formula (III):

Where:
R ₁ is selected from H, OH, CN, NH ₂ , or C _1-5 alkyl, C 1-5 heteroalkyl, C _2-5 alkynyl, C _3-6 cycloalkyl, and _3- to 6-membered heterocycloalkyl optionally substituted with 1, 2, or 3 R;
_R2 is selected from H, halogen, or _C1-3 alkyl and _C1-3 heteroalkyl optionally substituted with 1, 2 or 3 R;
m is selected from 0, 1, 2, 3, 4 and 5;
A is selected from phenyl or 5-6 membered heteroaryl optionally substituted with 1, 2 or 3 R;
R is selected from H, halogen, OH, CN, _NH2 , =O, _CH3 , _CH3CH2 , _CH3O , _CF3 , _CHF2 , _{CH2F ;}
The "hetero" in the C _1-5 heteroalkyl, 3- to 6-membered heterocycloalkyl, C _1-3 heteroalkyl, and 5- to 6-membered heteroaryl is each independently selected from N, -O-, =O, -S-, -NH-, -(C=O)-, -(S=O)-, and -(S=O) ₂ -;
In any of the above cases, the number of heteroatoms or heteroatom groups is each independently selected from 1, 2 or 3.

In some embodiments of the invention, in the above combination, the compound of formula (III) is selected from one of the following compounds:

In some embodiments of the invention, in the above combination, the compound of formula (III) is selected from one of the compounds of the following formulae:

好ましくは、前記Ｂ型肝炎表面抗原阻害剤は、下記の式（ＩＩＩｂ）で表される構造を有する。

Preferably, the Hepatitis B surface antigen inhibitor has a structure represented by formula (IIIa):

Preferably, the Hepatitis B surface antigen inhibitor has a structure represented by formula (IIIb):

In the combination described in the present invention, the reverse transcriptase inhibitor is selected from lamivudine, adefovir dipivoxil, entecavir, tenofovir disoproxil fumarate, or tenofovir alafenamide fumarate.

Preferably, the reverse transcriptase inhibitor is selected from entecavir or tenofovir disoproxil fumarate.

The combination described in the present invention is to prepare a pharmaceutical composition by mixing a compound of formula (I) or a pharma- ceutically acceptable salt thereof with a drug of any one of groups a to c as a medicament active ingredient.

Here, mixing the active pharmaceutical ingredients to produce a pharmaceutical composition means mixing two or three different drugs together as active pharmaceutical ingredients to produce a composite pharmaceutical composition.

The combination described in the present invention may be prepared by preparing a pharmaceutical composition using the compound of formula (I) or a pharma- ceutically acceptable salt thereof and a drug of any one of groups a to c as medicament active ingredients, and packaging the compositions separately so that they are taken separately.

Here, taking them separately includes taking one or two types first, then taking another one or two types, and taking two or three types simultaneously.

The use of the combination described in the present invention in the manufacture of a drug for the treatment of hepatitis B virus infection.

A pharmaceutical formulation composition according to the present invention, comprising the combination according to the present invention and at least one pharma- ceutically acceptable carrier and/or excipient.

The present invention further provides a kit comprising the combination described in the present invention or the pharmaceutical formulation composition described in the present invention.

The present invention further provides the use of the pharmaceutical composition or kit in the manufacture of a medicament for treating hepatitis B.

The method for packaging the combination of the present invention separately and administering it separately at the time of administration can be selected from the following methods.

[Administration Method]
The following is not intended to limit the methods of administration of the combinations of the present invention.

The components of the combination of the present invention can be formulated separately into a pharmaceutical composition, or some or all of them can be formulated together into a pharmaceutical composition. In some embodiments, the combination of the present invention can be formulated into a pharmaceutical composition suitable for single or multiple administration.

The components of the combination of the present invention may be administered individually or some or all of them may be administered together. The components of the combination of the present invention may not be administered substantially simultaneously or some or all of them may be administered substantially simultaneously. The components of the combination of the present invention may have the same or different administration cycles.

The components of the combinations of the invention can be administered independently by a variety of suitable routes, including, but not limited to, oral or parenteral administration (intravenous, intramuscular, subcutaneous, intraperitoneal, spinal or other parenteral routes, e.g., by injection or infusion). In some embodiments, the components of the combinations of the invention can be administered independently by oral or parenteral administration, e.g., intravenous or intraperitoneal injection.

The components of the combination of the present invention may each be independently in any suitable dosage form, including, but not limited to, tablets, troches, pills, capsules (e.g., hard capsules, soft capsules, enteric-coated capsules, microcapsules), elixirs, granules, syrups, injections (intramuscular, intravenous, intraperitoneal), granules, emulsions, suspensions, solutions, dispersions, and sustained release formulations for oral or parenteral administration.

Each component of the combination of the present invention may independently contain a pharma- ceutically acceptable carrier and/or excipient.

[Technical Effects]
The compound of formula (I) of the present invention is a hepatitis B core protein inhibitor, which can disrupt viral cccDNA library and inhibit HBV viral replication; the compound of formula (II) and formula (III) of the present invention is a hepatitis B surface antigen inhibitor, which can effectively reduce HBsAg. By combining the compound of formula (I) with said hepatitis B surface antigen inhibitor, or combining the compound of formula (I) with nucleoside or nucleotide reverse transcriptase inhibitor, or combining these three kinds of mechanism drugs, the replication process of HBV virus can be inhibited in a multi-channel manner, and the objectives such as improving therapeutic effect, reducing toxic side effects, shortening therapeutic cycle and dosage, reducing drug resistance, etc. can be achieved, and the effects of reducing HBV load, reducing HBsAg, and even removing can be achieved.

Definitions and Explanations
Unless otherwise stated, the following terms and phrases used herein have the following meanings: A particular term or phrase, unless otherwise defined, should be understood to have its ordinary definition, not to be indefinite or unclear. When a trade name appears in this specification, it refers to the corresponding product or its active ingredient.

As used herein, "pharmacologically acceptable" refers to those compounds, materials, compositions and/or dosage forms which are within the scope of sound medical judgment, suitable for contact with human and animal tissues, without significant toxicity, irritation, allergic response or other problem or complication, and consistent with a reasonable benefit/risk ratio.

The term "pharmaceutical acceptable salt" refers to a salt of a compound of the present invention, which is prepared with a relatively non-toxic acid or base, with the compounds having certain substituents discovered in this invention. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting these compounds with a sufficient amount of base, either alone or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting these compounds with a sufficient amount of acid, either alone or in a suitable inert solvent. Examples of pharma- ceutically acceptable acid addition salts include inorganic and organic acid salts, as well as salts of amino acids (e.g., arginine, etc.), and salts of organic acids such as glucuronic acid, such as inorganic acids including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, and the like, and organic acids including, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and similar acids. Some specific compounds of the present invention contain basic and acidic functional groups and can therefore be converted into any base or acid addition salt.

The pharma- ceutically acceptable salts of the present invention can be synthesized in a conventional manner from parent compounds that contain acid or basic groups. Typically, such salts are prepared by reacting these compounds in their free acid or base form with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both.

The term "pharmaceutical composition" refers to a mixture of one or more active ingredients herein or a pharmaceutical combination thereof with a pharmaceutically acceptable excipient. The purpose of a pharmaceutical composition is to facilitate administration of the compounds herein or pharmaceutical combinations thereof to a subject.

The term "pharmaceutical acceptable carrier" refers to any formulation or carrier medium, including water, oil, vegetable and mineral, cream bases, lotion bases, ointment bases, and the like, that can deliver an effective amount of an active agent of the present invention, does not interfere with the biological activity of the active agent, and has no toxic or adverse effects on the host or patient. These bases include suspending agents, tackifiers, penetration enhancers, and the like. Such formulations are well known to those skilled in the art of cosmetics or topical pharmaceuticals.

The term "excipient" generally refers to a carrier, diluent, and/or vehicle necessary to formulate an effective pharmaceutical composition.

The terms "include" and "including" are to be understood in an open and non-exclusive sense, i.e. "including but not limited to."

The term "treatment" means the administration of a compound or formulation described herein to prevent, ameliorate, or eliminate a disease or one or more symptoms associated with that disease, including:
(1) To prevent the appearance of a disease or disease state in a mammal, particularly where such a mammal is susceptible to the disease state but has not been diagnosed with the disease state;
(2) inhibiting a disease or disease state, i.e., arresting its progression;
(3) Alleviating the disease or disease state, i.e., causing the regression of the disease or disease state.

The term "effective amount" or "therapeutically effective amount" with respect to a pharmaceutical or pharmacologically active agent refers to a non-toxic but sufficient amount of the pharmaceutical or agent to achieve a desired effect. In the case of oral dosage forms of the present invention, an "effective amount" of one active agent in the composition refers to the amount necessary to achieve a desired effect when used in combination with another active agent in the composition. Determination of an effective amount varies from person to person and depends on the age and general condition of the subject, and also on the particular active agent, and the appropriate effective amount in each case can be determined by one of skill in the art based on routine experimentation.

The term "administration" refers to the physical introduction of a composition containing a therapeutic agent into a subject using any one of a variety of methods and delivery systems known to those skilled in the art. Routes of administration include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal or other parenteral routes, e.g., administration by injection or infusion. As used herein, the phrase "parenteral administration" refers to patterns of administration other than enteral and topical administration, typically by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcystic, subarachnoid, intraspinal, epidural and intrasternal injection and infusion, and in vivo electroporation. In some embodiments, the combination is administered by a non-parenteral route, and in some embodiments, is administered orally. Other non-parenteral routes include topical, epidermal or mucosal routes of administration, e.g., intranasal, vaginal, rectal, sublingual or topical administration. Administration can also be, for example, once, multiple times, and/or over one or more extended periods of time.

The term "subject" refers to a mammal. In some embodiments, the subject is a mouse. In some embodiments, the subject is a human.

As used herein, "combination" or "use in combination" means that two or more active agents can each be administered to a subject simultaneously in a single formulation, or each can be administered sequentially in any order in a single formulation.

The terms "active ingredient," "therapeutic agent," "active substance," or "active agent" refer to a chemical entity that is effective in treating a targeted disorder, disease, or condition.

The compounds of the present invention may exist in particular geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic and other mixtures thereof, such as mixtures enriched in enantiomers or non-enantiomers, and all such mixtures are included within the scope of the present invention. Other asymmetric carbon atoms may be present in substituents such as alkyl. All such isomers and mixtures thereof are both included within the scope of the present invention.

Unless otherwise stated, the terms "enantiomers" or "optical isomers" are stereoisomers that are mirror images of each other.

Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" refer to the inability to freely rotate around double bonds or single bonds of ring carbon atoms.

Unless otherwise explained, the term "diastereomers" refers to stereoisomers in which the molecules have two or more centers of chirality and are not mirror images of each other.

Unless otherwise stated, "(+)" means dextrorotatory, "(-)" means levorotatory, and "(±)" means racemic.

及び、
楔形点線結合

で一つの立体中心の絶対配置を、
棒状実線結合

及び、
棒状点線結合

で立体中心の相対配置を、
波線

で
楔形実線結合

又は、
楔形点線結合

を、或いは、
波線

で、
棒状実線結合

及び
棒状点線結合

を表す。 Solid wedge connections unless otherwise stated

And,
Wedge dotted line connection

The absolute configuration of one stereocenter is
Solid bar connection

And,
Bar dotted line connection

The relative configuration of the stereocenters is
Wavy line

Solid wedge connection at

Or,
Wedge dotted line connection

Or,
Wavy line

in,
Solid bar connection

and bar-dotted line bond

Represents.

The term "substituted" refers to the replacement of any one or more hydrogen atoms at a particular atom with a substituent, which may include deuterium and hydrogen variants, provided that the particular valence state is correct and the compound is stable after substitution. When the substituent is a keto group (i.e., =O), it means that two hydrogen atoms have been replaced. Keto group substitution does not occur in aromatic groups. The term "optionally substituted" refers to either substituted or unsubstituted, and unless otherwise defined, the type and number of substituents are any that are chemically stable and can be achieved.

When any variable (e.g., R) occurs more than once in a composition or structure of a compound, its definition is independent at each occurrence. So, for example, if a group is substituted with 0-2 R, then said group is optionally substituted with up to 2 R, and each occurrence of R is independently optional. Also, combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

When the number of linking groups is 0, for example, -(CRR) ₀ - means that the linking group is a single bond.

If one of the variables is a single bond, the two groups it connects are directly linked; for example, if L in A-L-Z is a single bond, the structure is actually A-Z.

The absence of a substituent indicates that the substituent is not present, for example, the absence of X in A-X indicates that the structure is actually A. If the atom through which a listed substituent is substituted is not specified, such a substituent may be bonded through any atom, for example, a pyridinyl group as a substituent may be bonded to the substituent through any carbon atom of the pyridine ring.

における連結基Ｌは－Ｍ－Ｗ－であり、この時－Ｍ－Ｗ－は左から右への読み取る順序と同じ方向に環Ａと環Ｂを構成

することができ、また、左から右への読み取る順序と逆方向に環Ａと環Ｂを構成

することもできる。上記連結基、置換基及び／又はその変形体の組み合わせは、このような組み合わせであれば安定した化合物になる場合のみ許容される。 If the listed linking group does not specify another linking direction, then the linking direction is arbitrary, for example:

In the formula, the linking group L is -M-W-, and in this case, -M-W- constitutes ring A and ring B in the same direction as the order of reading from left to right.

It is also possible to construct rings A and B in the reverse order of reading from left to right.

Combinations of the above linking groups, substituents and/or variables are permissible only if such combinations result in stable compounds.

直線破線結合

又は、
波線

で表すことができる。例えば、－ＯＣＨ_３の直線実線結合は、該基の酸素原子を介して他の基に結合されていることを意味する。

中の直線の破線結合は、該基内の窒素原子の両端が他の基に結合されていることを意味する。

中の波線は、当該フェニル基の部位１と２の炭素原子を介して他の基に結合されていることを意味する。

は、当該ピペリジニル基の任意の結合可能な部位が一つの化学結合によって他の基に結合できることを意味し、少なくとも

の四つの結合形態を含み、Ｈ原子が－Ｎ－に描かれていても、

には

この結合形態の基が含まれるが、一つの化学結合が接続されると、その部位のＨは一つ減少して対応する一価ピペリジン基になる。 Unless otherwise specified, when a group has one or more bondable sites, any one or more sites of the group can be bonded to other groups by chemical bonds. When the bonding mode of the chemical bond is delocalized and there is an H atom at the bondable site, the number of H atoms at the site is reduced to a group with a corresponding valence according to the number of bonded chemical bonds. The chemical bond by which the site is bonded to another group is:
Straight solid line connection

Straight dashed bond

Or,
Wavy line

For example, the straight solid bond of _-OCH3 means that the group is bonded to another group through the oxygen atom of that group.

A straight dashed bond within a group means that a nitrogen atom within the group is bonded at both ends to other groups.

The wavy line in the middle means that the phenyl group is bonded to another group via carbon atoms at positions 1 and 2.

means that any available bonding site of the piperidinyl group can be bonded to another group by one chemical bond, and at least

It includes the four bond forms, and even if the H atom is drawn as -N-,

for

Groups of this bond type are included, but when one chemical bond is connected, one H is removed at that site to form the corresponding monovalent piperidine group.

Unless otherwise stated, the number of atoms in a ring is generally defined as the number of ring members, e.g., a "5- to 7-membered ring" refers to a "ring" of 5 to 7 atoms arranged around it.

Unless otherwise defined, the term "C _1-3 alkyl" refers to a saturated hydrocarbon group made up of 1 to 3 carbon atoms in a straight or branched chain. The C _1-3 alkyl group includes C _1-2 and C _2-3 alkyl groups, which may be monovalent (e.g., methyl), divalent (e.g., methylene) and polyvalent (e.g., methine). Examples of _{C 1-3} alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), and propyl (including n-propyl and isopropyl). Unless otherwise defined, "C _2-8 alkenyl" refers to a saturated hydrocarbon group made up of 2 to 8 carbon atoms in a straight or branched chain, which includes at least one carbon-carbon double bond, and which may be located at any position in the group.

Unless otherwise defined, the term "4-6 membered heterocycloalkyl" by itself or in combination with other terms refers to a saturated cyclic group composed of 4 to 6 ring atoms, of which 1, 2, 3, and 4 ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, wherein the nitrogen atom is optionally quaternized and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) _p , where p is 1 or 2). It includes monocyclic and bicyclic ring systems, where bicyclic ring systems include spiro rings, fused rings, and bridged rings. Additionally, with respect to "4-6 membered heterocycloalkyl," a heteroatom can occupy the position at which the heterocycloalkyl is attached to the rest of the molecule. Said 4-6 membered heterocycloalkyl includes 5-6 membered, 4 membered, 5 membered, and 6 membered heterocycloalkyl, and the like. Illustrative examples of 4-6 membered heterocycloalkyl include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl (including tetrahydrothien-2-yl and tetrahydrothien-3-yl, and the like), tetrahydrofuranyl (including tetrahydrofuran-2-yl, and the like), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl, and 3-piperidinyl, and the like), piperazinyl (including 1-piperazinyl and 2-piperazinyl, and the like), morpholinyl (including 3-morpholinyl and 4-morpholinyl, and the like), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, or homopiperidinyl.

Unless otherwise stated, the following terms and phrases used herein have the following meanings. A particular phrase or term, unless specifically defined, should be understood to have its ordinary definition, not to be indefinite or unclear. When a trade name appears in this specification, it refers to the corresponding product or its active ingredient.

The intermediate compounds of the present invention can be prepared by various synthetic methods familiar to those skilled in the art, including the specific embodiments listed below, embodiments in combination with other chemical synthetic methods, and equivalent alternative methods familiar to those skilled in the art, and preferred embodiments include, but are not limited to, the examples of the present invention.

The chemical reactions of the specific embodiments of the present invention are completed in a suitable solvent, which should be suitable for the chemical reactions of the present invention and the reagents and materials required therefor. In order to obtain the compounds of the present invention, it may be necessary for those skilled in the art to modify or select synthetic steps or reaction schemes based on the existing embodiments.

The present invention will be described in detail below with reference to examples, but these examples do not limit the present invention in any way.

All solvents used in this invention are commercially available and may be used as is without further purification.

All solvents used in the present invention can be obtained from commercial sources. The present invention uses the following abbreviations: EtOH stands for ethanol, MeOH stands for methanol, TFA stands for trifluoroacetic acid, TsOH stands for p-toluenesulfonic acid, mp stands for melting point, EtSO ₃ H stands for ethanesulfonic acid, MeSO ₃ H stands for methanesulfonic acid, THF stands for tetrahydrofuran, EtOAc stands for ethyl acetate, THF stands for tetrahydrofuran, EA stands for ethyl acetate, DMAP stands for 4-dimethylaminopyridine, DCM stands for dichloromethane, and DIPEA stands for N,N-diisopropylethylamine.

FIG. 1 is a graph showing the effect of ex vivo combined administration of Compound 1 and TDF. FIG. 2 is a graph showing the therapeutic effect of test compounds on serum HBV DNA in AAV/HBV mice for 28 days. FIG. 3 is a graph showing the therapeutic effect of the test compound on the liver HBV DNA of AAV/HBV mice. FIG. 4 is a graph showing the therapeutic effect of test compounds on serum HBV RNA in AAV/HBV mice. FIG. 5 is a concentration profile of the test compound in the serum of AAV/HBV model mice. FIG. 6 shows changes in body weight of mice in each group.

The present invention will be described in detail below with reference to examples, but this is not intended to impose any adverse restrictions on the present invention. The present invention has been described in detail herein, and specific embodiments thereof have been disclosed. It is clear to those skilled in the art that various changes and modifications can be made in the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

Example 1 Preparation of Compound 1

Synthesis Route:

Step 1: Synthesis of Compound 1-A Anhydrous dichloromethane (5 L) was added to a dry 10 L three-neck flask, stirring was started, compound 1-SMA (500.00 g) and nitromethane were added to the three-neck flask in sequence, and the reaction system was placed in a dry ice ethanol bath and cooled to -10°C. The temperature was controlled at -10°C to 0°C, aluminum trichloride (1.15 kg) was slowly added to the reaction flask, the temperature was controlled below -0°C, α,α-dichlorodimethyl methyl ether (495.00 g) was slowly added to the reaction vessel, and the temperature was slowly raised to room temperature and stirred for 18 hours. TLC (PE:EA 3:1) monitoring showed that the raw material point disappeared and a new point with high polarity was generated. The reaction solution was withdrawn and slowly added dropwise to 10% potassium hydrogen sulfate solution (3 L), stirred for 20 minutes, and crushed ice was added to prevent overheating. The mixture was transferred to a 25 L separatory funnel and allowed to stand for separation, the dichloromethane layer was separated, and the aqueous phase was extracted with dichloromethane (2 L x 2). The organic phase was washed with 10% potassium hydrogen sulfate solution (5 L x 2), and the organic phase was separated and dried over anhydrous sodium sulfate (1 kg). The organic phase was concentrated under reduced pressure to obtain dark green solid compound 1-A.

¹ H NMR (400 MHz, deuterated chloroform) δ = 9.97 (br s, 1H), 9.87 - 9.82 (m, 1H), 7.58 (dd, J = 1.5, 3.3 Hz, 1H), 7.36 - 7.29 (m, 1H), 4.37 (q, J=7.1 Hz, 2H), 1.38 (t, J=7.2 Hz, 3H).

Step 2: Synthesis of Compound 1-B To a solution of compound 1-A (2 kg, 11.96 mol) in THF (20 L) was added p-toluenesulfonyl hydrazide (2.23 kg, 11.96 mol). Stirred at 20° C. for about 1 hour. After confirming the disappearance of the raw material by TLC, the reaction was heated to 60° C., then sodium cyanoborohydride (902 g, 14.36 mol) was added in batches, and after the addition was completed, the reaction was heated to 70° C. and stirred for 3 hours. After stopping heating and cooling to room temperature, the reaction was quenched by adding 5 L of water, most of the THC was removed under reduced pressure, and the residue was extracted with EA (1.5 L×3). The organic phases were combined, washed with saturated sodium chloride, and dried over anhydrous sodium sulfate. Filtration and removal of the solvent under reduced pressure gave the crude product as a pale yellow solid compound 1-B, which was separated by column chromatography.

Step 3: Synthesis of Compound 1-C Methanol (32 L) was added to a 50 L jacketed kettle, stirring was started, compound 1-SMB (4000.00 g) and diisopropylethylamine (5.25 L) were added in sequence, the internal temperature was lowered to 5-10°C, and benzyl mercaptan (2490.00 g) was slowly added dropwise to maintain the internal temperature at 5-15°C. After the addition was completed, the cooling system was closed, the temperature was allowed to rise naturally, and stirring was continued for 2.5 hours. Stirring was stopped, the rotation speed was adjusted to 100 rpm, the reaction liquid was discharged, and the filter was used to filter the cake, which was washed three times with water (5 L), then washed once with EtOH (3 L), and suction filtered until the cake was no longer viscous, to obtain pale yellow solid compound 1-C.

Step 4: Synthesis of Compound 1-D Dichloromethane (7.5 L) was added to a 50 L jacketed kettle, stirring was started, compound 1-C (1500 g) was added, the internal temperature was lowered to 0-10° C., and HCl solution (6 M, 4.12 L) was added. Under the condition of 0-10° C., sodium hypochlorite solution (commercially available 8% solution, 23.0 kg) was added dropwise through the opening, and after the addition was completed, the cooling system was closed and stirring was continued for about 17 hours through the opening. Then sodium hydrogen sulfite solution (1000 g, 5 L aqueous solution) was added dropwise thereto, and potassium iodide starch test paper was used to detect that no oxidant remained in the aqueous phase. Stirring was stopped, the mixture was allowed to stand and separated, the dichloromethane layer was collected, the aqueous layer was extracted with dichloromethane (2.5 L), and the dichloromethane layers were combined. The organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure to obtain a white solid compound 1-D.

¹ H NMR (400 MHz, deuterated chloroform) δ = 8.50 - 8.43 (m, 2H), 8.34 (d, J = 8.2 Hz, 1H), 4.04 (s, 3H).

Step 5: Synthesis of Compound 1-E Tetrahydrofuran (10 L) was added to a dry 50 L jacketed kettle, stirring was started, compound 1-B (2000 g) was added, and the internal temperature was lowered to 0-10° C. Within about 1.5 hours, potassium tert-butoxide (1 M THF solution, 15.67 L) was added while maintaining the temperature at 0-15° C., and after completion, the temperature was raised to about 20° C. and stirring was continued for 1 hour. The temperature was lowered to 0-10° C., and a solution of compound 1-D (4380 g) in tetrahydrofuran (10 L) was slowly added. After completion of the dropwise addition, the temperature was slowly raised to 15° C. and stirring was continued for about 16 hours. Ethyl acetate (10 L) was added for extraction, and the organic phase was washed twice with saturated sodium chloride solution (10 L), the aqueous phase was combined, extracted with EA (5 L), and the organic phase was combined. The organic phase was subjected to solvent removal under reduced pressure to obtain pale yellow solid compound 1-E.

^1H NMR (400MHz, DMSO- _d6 ) δ = 8.55 (d, J=1.4 Hz, 1H), 8.37 (dd, J=1.5, 8.3 Hz, 1H), 7.91 (d, J=8.3 Hz, 1H), 7.60 (s, 1H), 7.13 (d, J=1.8 Hz, 1H), 4.02 (q, J=7.0 Hz, 2H), 3.93 (s, 3H), 2.10 (s, 3H), 1.08 (t, J=7.1 Hz, 3H).

Step 6: Synthesis of Compound 1-SM1 Compound 1-E (1000.0 g) was added to a dry 10 L three-neck flask, stirring was started, glacial acetic acid (5 L) was added, and the reaction temperature was controlled at 25-30 ° C. Iron powder (1 eq, 140.9 g) was slowly added, and after stirring for 30 minutes, the second batch of iron powder (0.5 eq, 70.44 g) was slowly added, stirring was continued for 30 minutes, and then the third batch of iron powder (0.5 eq, 70.44 g) was slowly added, and after stirring for 30 minutes again, the fourth batch of iron powder (0.5 eq, 70.44 g) was added, and the reaction was continued with stirring until the raw material disappeared, and a new point with high polarity was generated. Stirring was stopped, and the reaction liquid was transferred to a 25 L separatory flask, 10 L ethyl acetate was added, and the mixture was washed with 5 L of saturated aqueous sodium hydrogen sulfate solution x 2, separated, and the aqueous phase was back-extracted with 5 L of ethyl acetate. The organic phases were combined and washed with 10% aqueous NaOH until pH>8, and the organic phase was separated and collected. The organic phase was concentrated under reduced pressure to give white solid compound 1-SM1.

^1H NMR (400MHz, DMSO- _d6 ) δ = 7.79 - 7.71 (m, 2H), 7.50 (d, J=1.8 Hz, 1H), 7.14 (dd, J=1.7, 8.5 Hz, 1H), 6.96 (d, J=2.0 Hz, 1H), 6.42 (s, 2H), 4.11 (q, J=7.2 Hz, 2H), 3.84 (s, 3H), 2.04 (s, 3H), 1.16 (t, J=7.1 Hz, 3H)

Step 7: Synthesis of Compound 1-F Toluene (12 L) was added to a dry 50 L jacketed kettle, stirring was started, 2-bromoethanol (9930 g) was added, followed by boron trifluoride ethyl etherate (268 g), and the reaction was allowed to warm to 30-35°C. Compound 1-SMC (3500 g) was slowly added dropwise, and the addition was completed in about 1.5 hours, at which time the internal temperature of the reaction was increased to about 55-65°C, the heater temperature was adjusted to 60°C, and the internal temperature was maintained at 55-65°C for 1 hour. The internal temperature of the reaction system was lowered to about 10°C, and an aqueous solution of sodium hydroxide (3783 g, 17.5 L of water) at about 20°C was slowly added into the reaction system, and the internal temperature was maintained at 10-20°C. After the NaOH solution was added, the temperature controller was stopped, and the reaction was allowed to continue stirring for about 16 hours. The stirring was stopped, allowed to settle, separated, the aqueous layer was extracted with 2-methyltetrahydrofuran (10 L), the organic phases were combined, washed with water (10 L), allowed to settle, separated, and the organic phase was collected and concentrated under reduced pressure to give compound 1-F as a colorless oil.

^1H NMR (400MHz, deuterated chloroform) δ = 3.87 - 3.71 (m, 4H), 3.66 - 3.59 (m, 3H), 3.42 (dd, J = 6.0, 11.7 Hz, 1H), 3.20 - 3.13 (m, 1H), 2.79 (t, J=4.6 Hz, 1H), 2.65 - 2.59 (m, 1H)

Step 8: Synthesis of Compound 1-G Aqueous sodium hydroxide (3240 g, 15 L water) was added to a 50 L jacketed kettle, compound 1-F (4430 g) was added, heating was started, and the reaction was allowed to warm to 90° C. and then stirred for 1 hour. Cooling was started, lowered to about 15° C., a solution of p-toluenesulfonyl chloride in tetrahydrofuran (6180 g, 15 L tetrahydrofuran) was added, the temperature controller was closed, and the reaction was allowed to stir at about 15° C. for about 16 hours. Stirring was stopped, allowed to settle, separated, the aqueous phase was extracted with 2-methyltetrahydrofuran (10 L), the 2-methyltetrahydrofuran phase (contained white insoluble matter, which disappeared after washing with water) was washed with water (5 L), and the organic phase was combined. DMAP (500 g), triethylamine (2.5 L) were added to the organic phase, stirred for 30 minutes, saturated sodium chloride solution (10 L) was added to wash, allowed to settle, separated, and the aqueous phase was discarded. The organic phase was washed with potassium hydrogen sulfate solution (3800 g, water 15 L) and saturated sodium chloride solution (5 L x 2), and then allowed to stand and separate, and the organic phase was collected. The organic phase was concentrated under reduced pressure to remove the solvent, and the crude product compound 1-G was obtained.

¹ H NMR (400 MHz, deuterated chloroform) δ = 7.77 (d, J=8.3 Hz, 2H), 7.34 (d, J=8.2 Hz, 2H), 4.03 - 3.91 (m, 2H), 3.80 - 3.49 (m, 8H), 3.33 (dd, J=9.9, 11.4 Hz, 1H), 2.43 (s, 3H)

Step 9: Synthesis of Compound 1-H Acetone (30 L) was added to a clean 50 L jacketed kettle, stirring was started, Compound 1-G (4500 g) was added, followed by sodium iodide (6190 g), heating was started, and the reaction was allowed to warm to 75° C. and then stirred for 16 hours. After cooling to room temperature, it was filtered and the filtrate was concentrated under reduced pressure at 50° C. Ethyl acetate (15 L) and water (10 L) were added to the concentrated crude product, stirred, allowed to stand and separated, and the organic phase was washed with 0.5 M sodium thiosulfate (10 L). The aqueous phase and sodium thiosulfate solution were combined and then extracted with EtOAc (5 L). The organic phases were combined and washed with saturated sodium chloride solution (10 L), allowed to stand and separated, and the organic phase was collected. The organic phase was concentrated under reduced pressure to remove the solvent and give the crude product Compound 1-H.

^1H NMR (400MHz, deuterated chloroform) δ = 3.90 - 3.83 (m, 2H), 3.81 - 3.75 (m, 2H), 3.74 - 3.65 (m, 5H), 3.63 - 3.49 (m, 7H), 3.31 - 3.18 (m, 3H), 3.06 - 3.04 (m, 2H)

Step 10: Synthesis of Compound 1-I DMSO (20 L) was added to a clean 50 L jacketed kettle, stirring was started, compound 1-H (4700 g) was added, the temperature was raised to 35°C, then sodium cyanide (1010 g) was added, the internal temperature of the reaction was raised to about 60°C within 20 minutes, then gradually lowered to 35°C, and stirring was continued for about 16 hours. Sodium bicarbonate solution (2000 g sodium bicarbonate, 10 L water) was added to the reaction system, stirring was continued for about 5 minutes, EtOAc:MeOH (20 L, 2 L) was added, stirring was continued for 2 minutes, and the mixture was allowed to stand for about 1 hour. The lower layer solution was separated, about 30 L, and extracted twice with EtOAc:MeOH (15 L:1.5 L for the first time, and 5 L:0.5 L for the second time). The upper organic phase after extraction and the remaining upper layer of the reaction solution were combined, washed three times with saturated sodium chloride solution (10 L each), allowed to stand, separated, the aqueous phase was discarded, and the organic phase was collected. The organic phase was decompressed to remove the solvent, and the crude product was separated by column chromatography to obtain a colorless oily substance, Compound 1-I.

¹ H NMR (400 MHz, deuterated chloroform) δ = 3.84 - 3.65 (m, 6H), 3.61 - 3.53 (m, 2H), 3.35 (t, J = 10.5 Hz, 1H), 2.49 - 2.44 (m, 2H).

Step 11: Synthesis of Compound 1-J Under the protection of argon gas, Raney Nickel (10.00 g, 116.73 mmol) and EtOH (150 mL) were added to a dry hydrogenation flask, followed by 1-I (20 g, 157.31 mmol) and NH ₃ .H ₂ O (13.65 g, 97.36 mmol, 15.00 mL, 25% purity), which was then purged and the reaction was stirred at 50 psi and 50° C. for 3.5 hours. The reaction was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to give a yellow oily compound 1-J.

^1H NMR (400MHz, deuterated chloroform) δ = 3.82 - 3.57 (m, 6H), 3.34 - 3.18 (m, 1H), 2.86 - 2.72 (m, 2H), 1.60 - 1.38 (m, 2H).

Step 12: Synthesis of compound 1-SM2 1-J (800.00 g) was added to a 5 L three-neck flask, stirring was started, ethyl acetate (800 mL) was added within about 0.5 h, HCl/EtOAc (1.6 L) 4M was slowly added dropwise until the reaction pH was <5, and the internal temperature was maintained at 5-15° C. The cooling system was closed, the temperature was raised to room temperature, and stirring was continued for 1 h. Stirring was stopped and filtered through a tabletop filter to obtain a cake. The cake was concentrated under reduced pressure (40-45° C.) to obtain the crude product. Acetonitrile (2 mL/g) was added to the above product and slurried for 1 h. The cake was collected by filtration through a tabletop filter, and the organic solution was removed under reduced pressure to obtain a white solid compound 1-SM2.

^1H NMR (400MHz, _CD3OD ) δ = 3.88 - 3.72 (m, 5H), 3.67 - 3.59 (m, 1H), 3.36 -3.31 (m, 1H), 3.14 (t, J = 6.7 Hz, 2H), 1.87 - 1.67 (m, 2H).

Step 13: Synthesis of Compound 1-1-A and Compound 1-1-B Toluene (20 L) was added to a dry 50 L jacketed kettle, stirring was started, Compound 1-SM1 (2500 g) was added, and the internal temperature was raised to 30-35°C. Nitrogen gas was purged to maintain an inert gas atmosphere inside the kettle, and trimethylaluminum (3.0 L, the internal temperature of the kettle increased slowly due to the addition of Al(CH ₃ ) ₃ ) was added dropwise, and after completion, the nitrogen gas was closed, the temperature was raised to 80-85°C, and stirring was continued for about 16 hours. Cooling was started, the reaction temperature was lowered to 20-30°C, and about half of the reaction liquid, about 12 L, was transferred, and EToAc (10 L) was added and mixed uniformly. The mixture was added to 10% KHSO ₄ solution (10 L) while stirring, stirred for 2 minutes, allowed to settle, separated, and the organic layer was washed with additional 10% KHSO ₄ solution (10 L), and the aqueous phases were combined and extracted twice with DCM (7.5 L each). The remaining half of the reaction solution, about 12 L, was transferred and treated in the same manner as above, and the organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was then slurried with two volumes of n-heptane for 1 hour. It was filtered and dried under vacuum (>12 hours, temperature 40°C, P≦-0.1 MPa). A mixture of compound 1-1-A and compound 1-1-B was obtained.

Step 14: Synthesis of Compound 1-2 Tetrahydrofuran (3840 mL) was added to a 10 L three-neck flask, stirring was started, a mixture of Compound 1-1-A and Compound 1-1-B (480.00 g) was added, and a solution of LiOH.H ₂ O (118.84 g) in H ₂ O (960 mL) was slowly added dropwise. After the dropwise addition was completed, the temperature was raised to 60° C., and stirring was continued for 1 hour. Concentrated HCl was added to the reaction solution to adjust the reaction system pH to 2, and stirring was stopped. The mixture was allowed to stand and separated. The aqueous phase was extracted twice with THF (600 mL), and the organic phase was combined. The organic phase was concentrated under reduced pressure (40-45° C.), the solid was slurried with pure water (2 mL/g) for 0.5 hours, filtered, and the cake was dried in vacuum (>12 hours, temperature 40° C., P≦−0.1 MPa) to obtain Compound 1-2.

^1H NMR (400MHz, DMSO-d6) δ = 11.19 (s, 1H), 8.09 (d, J=8.2 Hz, 1H), 8.00 (d, J=1.6 Hz, 1H), 7.88 (dd, J=1.5, 8.3 Hz, 1H), 7.39 (dd, J=1.2, 2.0 Hz, 1H), 7.01 (d, J=2.0 Hz, 1H), 2.05 (s, 3H).

Step 15: Synthesis of Compound 1 DMF (2.25 L) was added to a 5 L three-neck flask, and stirring was started. Compound 1-2 (400.00 g) and HATU (744.83 g) were added in sequence and stirred for 30 minutes, and then compound 1-SM2 (229.86 g) was added. Within 1 hour, DIPEA (568.68 mL) was slowly added dropwise at room temperature, and after the dropwise addition was completed, the mixture was stirred at room temperature and stirred for 16 hours. The reaction solution was transferred to a separatory funnel, and ethyl acetate (2 L) and pure water (1 L) were added and stirred for 2 minutes. The mixture was left to stand, and the aqueous phase was separated. Further pure water (1 L) was added to wash, stirred, left to stand, and separated. The combined aqueous phase was extracted three times with EtOAc (500 mL), and the organic phase was combined. The organic phase was washed successively with sodium carbonate solution (1.5 L) twice, potassium hydrogen sulfate solution (1 L) twice, and purified water (1 L) twice. The organic phase was concentrated under reduced pressure (40-45° C.) to obtain a crude product. Ethyl acetate (2 mL/g) was added to the crude product and slurried for 1 hour. The cake was collected by filtration to obtain compound 1.

^1H NMR (400MHz, DMSO- _d6 ) δ = 11.13 (br s, 1H), 8.73 (br t, J = 5.5 Hz, 1H), 8.05 (d, J = 8.2 Hz, 1H), 7.83 (d, J =1.3 Hz, 1H), 7.74 (dd, J =1.5, 8.4 Hz, 1H), 7.36 (s, 1H), 6.98 (d, J =2.0 Hz, 1H), 3.71 - 3.48 (m, 5H), 3.45 - 3.31 (m, 1H), 3.45 - 3.30 (m, 1H), 3.27 - 3.21 (m, 1H), 3.14 (dd, J = 9.9, 11.2 Hz, 1H), 2.03 (s, 3H), 1.53 (q, J = 7.0 Hz, 2H).

Biological test experiments

Experimental Example 1: Study of ex vivo coadministration of Compound 1 and Tenofovir disoproxil fumarate (TDF) on HBV inhibitory activity

1. Experimental Method 1.1 On day 1, HepG2.2.15 cells were seeded into 96-well cell culture plates at a density of 40,000 cells/well, and then the cells were cultured overnight at 37° C. with 5% CO ₂ .

1.2 On the second day, compound 1 and TDF were orthogonally distributed at seven different concentrations (selected at a concentration gradient of approximately 8x, 4x, 2x, 1x, 0.5x, 0.25x, and 0.125x _EC50 ) and added to a 96-well plate, with three duplicate wells for each combination, and the final concentration of DMSO was 0.5%. The detection concentrations of each compound are shown in Table 1.

1.3 On day 5, the cell supernatant was discarded, new medium containing the compound was added, and the cells were cultured for 3 days at 5% CO ₂ and 37° C. On day 8, DNA was extracted from the supernatant of the cell plate after compound treatment according to the instructions of the QIAamp 96 DNA Blood Kit (12).

1.4 HBV DNA was quantified by qPCR. HBV plasmid DNA was used as the standard, and the standard HBV plasmid DNA concentration was diluted 7 times in a 10-fold gradient from ¹⁰⁷ copies/μL. A standard curve was fitted using the HBV DNA copy number and CT value of each standard, and the HBV DNA copy number in each test sample was calculated.

The HBV DNA copy number test data was processed using MacSynegy software (Pricard et al., 1990) to analyze the effect parameters of the combined administration of compound 1 and TDF. The cytotoxicity of the compounds against HepG2.2.15 cells was detected using a CellTiter-Glo kit. The chemiluminescence intensity (RLU) of each cell well was detected using a multimode microplate reader according to the method in the kit's instructions.

2. Results The HBV inhibitory activity of compound 1 and TDF administered in vitro was evaluated using HepG 2.2.15 cells. The results of the combined administration of drugs on HBV inhibitory activity and cytotoxicity are summarized in Table 2, and the effect diagram of the combined administration is shown in Figure 1.

The results of the experiment showed that the synergy index and antagonism index in the 95% confidence space of the in vitro coadministration of compound 1 and TDF were 13.27 and -1.74, respectively, indicating a positive effect. None of the compounds showed cytotoxicity within the tested concentrations (see Table 3).

Note:
Description of the overall index of drug combination: if the absolute value of the index is < 25, that is, it is a positive effect; if the absolute value of the index is in the range of 25-50, that is, it is a mild but obvious synergistic or antagonistic effect; if the absolute value of the index is in the range of 50-100, that is, it is a moderate synergistic or antagonistic effect, which may have significant significance to the body's actions; if the absolute value of the index is > 100, that is, it is a high degree of synergistic or antagonistic effect, which may have significant significance to the body's actions.

3. Conclusions The in vitro combination administration of compound 1 and TDF showed a positive effect on the inhibitory activity of HBV, and the compound showed no cytotoxicity within the tested concentrations. The test results support the clinical combination of compound 1 and TDF in the treatment of patients with chronic HBV infection.

Experimental Example 2: Evaluating the in vivo anti-Hepatitis B virus efficacy of test compounds using an AAV/HBV mouse model

Experimental materials

1. Animals Five-week-old specific pathogen-free male C57BL/6 mice were purchased from SHANGHAI SLAC LABORATORY ANIMAL CO. LTD.

2. Solvents and Compound Solvents:
10% Solutol solution

Test Compound:
After adding an appropriate amount of compound 1 to the above solvent and vortexing, a uniform suspension of particles was obtained, and the concentrations prepared were 1.0, 3.0 and 10.0 mg/mL. Stored at 4°C until use. Compound 1 was calculated according to a salt coefficient of 1.0 and a purity of 100% when prepared.
Tenofovir disoproxil fumarate (TDF) was purchased from Shanghai Panhong Chemical Technology Co., Ltd. An appropriate amount of TDF was weighed and dissolved in saline to prepare a 1 mg/mL mother solution, which was then vortexed until the TDF was completely dissolved, dispensed into 1 mL portions, and stored at -20°C. Before each administration, 1 mL of the mother solution was collected and diluted 10-fold with saline to a working solution of 0.1 mg/mL, which was then used for administration on the day.

Recombinant virus rAAV8-1.3HBV
rAAV8-1.3HBV (type D, ayw) was purchased from Beijing FivePlus Molecular Medicine Institute Co. Ltd., 5+MMI, with a batch number of x2018032301, and 1×10 ¹² viral genome (v.g.)/mL. It was diluted to 5×10 ¹¹ v.g./mL with sterile PBS before the experiment. Each mouse was injected with 200 μL, i.e., each mouse was injected with 1×10 ¹¹ v.g.

3. Test Methods Construction of AAV/HBV Mouse Model AAV/HBV Injection: rAAV8-1.3HBV was prepared in advance using sterile PBS to give a solution with a concentration of 1×10 ¹¹ vg/200 μL prior to injection.

Infection level detection:
On days 14 and 21 after virus injection, ∼120 μL of blood was collected from the submandibular vein of all infected mice for serum collection. Whole blood was incubated in a 37°C incubator for 30 min, and then centrifuged at 13,200 × g at 4°C for 3 min to collect ∼30 μL of serum. Serum was stored at −80°C and used for detection of HBV DNA, HBeAg, and HBsAg.

Grouping:
On day 28 after virus injection, groups were divided based on HBV DNA, HBsAg and HBeAg levels in serum samples on days 14 and 21 after virus injection, and on mouse body weight.

Experimental date definition:
The day of the first administration was designated as day 0 of the experiment.

4. In vivo Experimental Design The dosing and sampling scheme for the in vivo experiment is as shown in Table 4.

Serum sample preparation:
Blood samples were incubated at 37° C. for .about.30 min, then centrifuged at 13,200 g for 3 min at 4° C., and the separated supernatant was flash frozen on dry ice.

Plasma sample preparation and pretreatment:
Blood samples were anticoagulated with _K2EDTA and then centrifuged at 7,000g for 10 min at 4°C to separate the supernatant. The separated plasma was mixed with a precipitant in a ratio of 1:20 [methanol:acetonitrile (v:v, 50:50) solution], vortexed, and flash frozen on dry ice.

Weight Recording:
During the course of the in vivo experiment, the condition of the mice was regularly observed, and the body weights of the mice were recorded on the days of infection, administration, blood sampling, and the termination of the experiment.

Serum was collected by cardiac blood sampling and used to detect HBV DNA and HBV RNA levels in groups 1 and 4.

5. Sample Analysis 1. HBsAg ELISA (Antu Bio, CL 0310) kit instructions were used to detect the HBsAg content in mouse serum.

2. HBeAg content in mouse serum was detected using the HBeAg ELISA kit (Antu Bio, CL 0312) instructions.

3. HBV DNA content in mouse serum and liver was detected by quantitative PCR.

Data Analysis:
Data are expressed as the mean ± standard error of the samples in each group, unless otherwise stated, Groups 1-6: n = 8. Statistical analysis was performed using Student's t test.

6. Results 1) Effect of test compound on serum HBV DNA in AAV/HBV mouse experiment The HBV DNA content in the serum of mice in each group is summarized in FIG.
The serum HBV DNA content of mice in the vehicle group (Group 1) remained stable after administration, and the compound 1 single agent group: compared with the vehicle group (Group 1), the HBV DNA content in the serum of mice in the three dose groups of compound 1, low (10 mpk, Group 2), medium (30 mpk), and high (100 mpk), began to show a dose-dependent decrease three days after administration, the HBV DNA content in the low and medium dose groups remained stable seven days after administration, and the HBV DNA content in the serum of the high dose group continued to decrease from 7 to 28 days after administration, and was significantly lower than the vehicle group (p<0.01).

Compared to the vehicle group (Group 1), the HBV DNA content in mouse serum decreased 3 days after administration, and the HBV DNA content in serum remained stable for 7 to 28 days after administration. Compared to the compound 1 (Group 2, 10 mpk) and TDF (Group 5, 1 mpk) single agent groups, the combination administration showed a significant effect of lowering the HBV DNA content in mouse serum (p<0.01).

2) Effect of test compound on HBV DNA in liver in AAV/HBV mouse experiment The HBV DNA content in the liver of mice in each group is summarized in FIG.
Compared with the vehicle group (Group 1), after 28 days of administration, the HBV DNA content in the liver of mice in the three dose groups of Compound 1, low (10 mpk), medium (30 mpk), and high (100 mpk), was reduced. After 28 days of administration, the HBV DNA content in the liver of the group administered in combination with Compound 1 and TDF (Group 6, 10+1 mpk) was significantly lower than that of the group administered with Compound 1 alone at 10 mpk (p<0.001).

3) Effect of test compound on serum HBV RNA in AAV/HBV mouse experiment The HBV RNA content in mouse serum of each group is summarized in FIG.
Compared with the vehicle group (Group 1), HBV RNA content in mouse serum from the high dose (100 mpk) group of Compound 1 was reduced 28 days after administration.

4) Pharmacokinetic analysis of test compound in AAV/HBV model mice The drug concentrations in the serum of mice in Group 2 are summarized in FIG.
On day 27, the mean drug concentrations in mouse serum at 0 h (pre-dose), 1, 4, 8, 9, 12, and 24 h after the first dose were 78, 122,200, 1630, 605, 10,300, 1280, and 66 nM, respectively. Drug absorption peaked 1 h after dosing, with a plasma half-life of 2.23 h and an AUC _0-inf of 47,600 nM.h.

5) Monitoring of Mouse Health and Weight The health and weight of the mice were monitored regularly throughout the experiment.
The results of changes in mouse body weight, compared with the body weight on day 0, are summarized in FIG.
During the administration period, the body weight of all mice was stable and did not decrease significantly, and the condition of the mice was good.

Note:
The changes in body weight of the mice were recorded from day -28 to day 28 (the day of the first administration was designated as day 0). The body weight on day 0 was used as a baseline for comparison, and a 20% weight loss was defined as a humane endpoint according to IACUC regulations, and mice that lost 20% or more of their body weight had to be removed from the study.

7. Conclusion Compared with the solvent control group, the serum and liver HBV DNA of mice treated with compound 1 was significantly reduced, and showed a dose-dependent trend in serum. After the combination of compound 1 with TDF, the effect was significantly better than that of compound 1 alone, showing good combination administration effect. During the experimental period, the body weight of the mice did not decrease significantly, indicating that the mice tolerated the test compounds well.

8. Description: In the above experiment, TDF was replaced with ETV (entecavir), and the combined use of compound 1 and ETV achieved an in vivo antiviral effect, which significantly reduced mouse serum and liver HBV DNA, which was superior to the treatment with compound 1 alone.

Experimental Example 3: Study on HBV inhibitory activity of in vitro combination of Compound 1 with the following compounds of formula (IIb) and formula (IIIb)

1. Experimental material Cell lines:
HepG2.2.15 cells were constructed and provided by WuXi AppTec, and the cell culture medium was DMEM/F12 medium supplemented with 2% fetal bovine serum, 2 mM glutamine, 1× non-essential amino acids, 100 U/mL penicillin, and 100 μg/mL streptomycin.

reagent:
The main reagents used in this study included FastStart Universal Probe MasterMaster (Roche, Catalog No. 04914058001), DNA Extraction Kit (Qiagen, Catalog No.: 51162), and CellTiter-Glo (Promega-G7573).

device:
The main equipment used in this study was a 7900 real-time PCR instrument (Applied Biosystems), a multifunction microplate reader (BioTek, Synergy2), and a QuantStudio ^™ 6Flex System (Applied Biosystems).

Test Compound:
Compound 1, compound of formula (IIb), compound of formula (IIIb).

2. Experimental method

2. Anti-HBV activity of single compounds

2.1.1 On day 1, HepG2.2.15 cells were seeded in a 96-well cell culture plate at a density of 60,000 cells/well, and then the cells were cultured overnight at 5% CO ₂ and 37° C. On day 2, compound 1, compound of formula (IIb), and compound of formula (IIIb) were diluted and added to a 96-well plate, and three duplicate wells were set for each combination, and the cells were cultured at 5% CO ₂ and 37° C. for 3 days. The detection concentration of each compound is shown in Table 7. On day 5, the medium was replaced with a new medium containing the compound, and on day 8, the supernatant was collected. HBsAg in the supernatant was detected by ELISA, and at the same time, DNA in the supernatant was extracted, and the content of HBV DNA in the supernatant was measured by quantitative PCR. After collecting the cell supernatant, CellTiter-Glo was added to detect the viability of the cells.

2. Anti-HBV activity of two-compound combinations

2.2.1 On day 1, HepG2.2.15 cells were seeded in a 96-well cell culture plate at a density of 60,000 cells/well, and then the cells were cultured overnight under the conditions of 5% _CO2 and 37°C. On day 2, different concentrations of compounds were added to treat the cells. The compounds were diluted in a gradient and measured in parallel in a combination of 7x7 concentrations in triplicate wells. The compound concentrations in the combination activity test were based on the _EC50 value of each compound, and the seven concentration gradients were about 2x, 1x, 1/2x, 1/4x, 1/8x, 1/16x and 1/ _32xEC50 , respectively. The compound concentrations in the combination activity test experiment are shown in Table 8, and the configuration of combination administration is shown in Table 9. On day 5, the medium was replaced with fresh medium containing the compounds. On day 8, the culture supernatant was collected, DNA was extracted, and the content of HBV DNA was measured by quantitative PCR. At the same time, cell viability was detected with CellTiter Glo reagent, and the combined effect of the two drugs was analyzed using MacSynergy software.

Note:
a=compound 1, b=compound of formula (IIb) or compound of formula (IIIb), 0 represents no compound, 1-7 represent 7 concentrations.

3. Experimental results:
The inhibitory activity of the compound 1 and the compound of formula (IIb) and the compound of formula (IIIb) administered in combination against HBV in vitro was evaluated using HepG2.2.15 cells. The inhibitory activity and cytotoxicity of the single compound against HBV are summarized in Table 10, and the inhibitory activity and cytotoxicity of the combined administration against HBV are summarized in Tables 11 to 13.

Note:
Description of the drug combination index: if the absolute value of the index is <25, that is, it is a positive effect; if the absolute value of the index is in the range of 25-50, that is, it shows a mild but obvious synergistic or antagonistic effect; if the absolute value of the index is in the range of 50-100, that is, it shows a moderate synergistic or antagonistic effect, which may have significant significance on the effect in the body; if the absolute value of the index is >100, that is, it shows a high degree of synergistic or antagonistic effect, which may have significant significance on the effect in the body.

4. Experimental conclusion In the HepG2.2.15 in vitro HBV infection model, the test compounds inhibited HBV DNA in a dose-dependent manner, and the combination of compound 1 and the compound of formula (IIb) and the compound of compound 1 and the compound of formula (IIIb) showed positive results and did not show cytotoxicity within the combination test concentration. The test results support the clinical combination of compound 1 and the hepatitis B surface antigen inhibitors such as the compound of formula (IIb) and the compound of formula (IIIb) in the treatment of chronic HBV infection.

又は、化合物３

に置き換えて、化合物２と式（ＩＩｂ）の化合物、式（ＩＩＩｂ）の化合物との併用投与結果、及び化合物３と式（ＩＩｂ）の化合物、式（ＩＩＩｂ）の化合物との併用投与結果を得、いずれも効果がプラスするデータ結果を示し、かつ併用試験濃度内では細胞毒性を示さなかった。試験結果は、慢性ＨＢＶ感染の治療における化合物２、化合物３などの当該類化合物が前記式（ＩＩｂ）の化合物、式（ＩＩＩｂ）の化合物などのＢ型肝炎表面抗原阻害剤の、臨床的に併用を支持した。 5. Description According to the above embodiment, compound 1 is converted to compound 2.

Or, Compound 3

The results of the combined administration of compound 2 with the compound of formula (IIb) and the compound of formula (IIIb), and the combined administration of compound 3 with the compound of formula (IIb) and the compound of formula (IIIb) were obtained, and all of the data showed positive effects and no cytotoxicity was observed within the combined test concentrations. The test results support the clinical combination of the compounds of the same class, such as compound 2 and compound 3, with the hepatitis B surface antigen inhibitors, such as the compound of formula (IIb) and the compound of formula (IIIb), in the treatment of chronic HBV infection.

Claims (6)

Compounds of formula (I)

or a pharma- ceutically acceptable salt thereof , and a compound selected from the group a to c below:
a. Hepatitis B surface antigen inhibitors;
b. reverse transcriptase inhibitors;
c. Hepatitis B surface antigen inhibitors and reverse transcriptase inhibitors;
and any one of the following pharmaceutical compositions.
(wherein the compound of formula (I) is the following compound

It is one of the
The hepatitis B surface antigen inhibitor is

It is one of the
The reverse transcriptase inhibitor is selected from lamivudine, adefovir dipivoxil, entecavir, tenofovir disoproxil fumarate, or tenofovir alafenamide fumarate. 2. The composition of claim 1, wherein the reverse transcriptase inhibitor is selected from entecavir and tenofovir disoproxil fumarate. A medicament for the treatment of Hepatitis B virus infection comprising the composition of claim 1 . A pharmaceutical composition comprising the composition of claim 1 and at least one pharma- ceutically acceptable carrier and/or excipient. A kit comprising the composition according to claim 1 or the composition according to claim 4 . A medicament for treating hepatitis B, comprising the composition according to claim 4 or the kit according to claim 5 .

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